Grant: Alliance Grants

Understanding the biological processes involved in viral infection of SARS-CoV-2 is required to effectively treat and prevent COVID-19 outbreaks and potential future coronavirus pandemics. Elucidating the biology of virus replication cycles and its evolution begins with sequencing of its genomic RNA. This facilitate the identification of key proteins involved and mutations as the COVID-19 pandemic develops. The RNA sequence of a virus only tells part of the story - methylation of viral RNA within infected host cells has been recently shown to alter synthesis and expression of viral proteins and enable viruses to "disguise" themselves to evade the host's immune system. Determining the location of these methylation sites in RNA is critical to guiding future research to tackle COVID pandemics; however, current methods to identify these locations quantitatively and at the single-nucleotides level in a high-throughput manner is lacking. The proposed sequencing method will implement a highly-selective chemical process that is compatible with currently existing high-throughput sequencing systems and readily identifies the presence and level of methylation at the N6 -position of adenosine - the most prevalent methylation form. To achieve the goals of this project, we are partnering with Princess Margaret Cancer Centre and its BioBank in Toronto which has developed a repository of COVID-19+ patient samples. Together, the team provides state-of-the-art genomics expertise and facilities to develop the proposed technology and map the genomic methylation landscape of SARS-CoV-2 RNA in infected human samples. Our one-year research plan is feasible and expected to lead to the development of a robust sequencing platform to be used in the genomic centres to facilitate research in the biology of RNA methylation in coronavirus infection in humans.